Classifications

• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
  • A61Q19/00—Preparations for care of the skin
• • A—HUMAN NECESSITIES
  • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
  • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
  • A23L7/00—Cereal-derived products; Malt products; Preparation or treatment thereof
  • A23L7/10—Cereal-derived products
  • A23L7/104—Fermentation of farinaceous cereal or cereal material; Addition of enzymes or microorganisms
  • A23L7/107—Addition or treatment with enzymes not combined with fermentation with microorganisms
• • A—HUMAN NECESSITIES
  • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
  • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
  • A23L7/00—Cereal-derived products; Malt products; Preparation or treatment thereof
  • A23L7/10—Cereal-derived products
  • A23L7/115—Cereal fibre products, e.g. bran, husk
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K8/00—Cosmetics or similar toiletry preparations
  • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
  • A61K8/72—Cosmetics or similar toiletry preparations characterised by the composition containing organic macromolecular compounds
  • A61K8/73—Polysaccharides
• • C—CHEMISTRY; METALLURGY
  • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
  • C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
  • C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
  • C08B37/0006—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid
  • C08B37/0024—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid beta-D-Glucans; (beta-1,3)-D-Glucans, e.g. paramylon, coriolan, sclerotan, pachyman, callose, scleroglucan, schizophyllan, laminaran, lentinan or curdlan; (beta-1,6)-D-Glucans, e.g. pustulan; (beta-1,4)-D-Glucans; (beta-1,3)(beta-1,4)-D-Glucans, e.g. lichenan; Derivatives thereof

Definitions

• Soluble dietary fibre from oat and barley grains method for producing a fraction rich in B-glucan and use of the fraction in foods, pharmaceuticals and cosmetics. . DESCRIPTION
• the present invention relates to a process for the cost-effective extraction of high molecular weight soluble dietary fibres and functional non-starch polysaccharides, from oat and barley grains and the downstream enrichment and utilisation of these materials.
• a novel methodology to produce ⁇ -glucans of high and medium molecular weight, in a controlled, cost effective manner, is described.
• soluble dietary fibres present in oat and barley are not digested in the human intestine and therefore pass through to the colon where they are available for microbial fermentation and as such are effective prebiotic materials.
• the soluble ⁇ -glucans from oat and barley are very interesting as functional ingredients in foods as they exhibit gelling behaviour, stabilising properties, water binding and impart good mouth feel to products.
• High molecular weight ⁇ -glucans have potential as viscosity modifiers, colloidal stabilisers, texturisers etc in foodstuffs.
• Inglett in two patent applications (US 4,996,063 and WO 92/10106) describes methods to produce water-soluble dietary fibre compositions from milled, heat -treated oat flours and milled barley flours, via treatment with ⁇ -amylase enzymes to degrade starch components and subsequent centrifugation to remove insoluble materials from the hydrolysate mixture.
• the products are relatively low in soluble dietary fibre content, with no reference to the molecular weight of the ⁇ -glucan components. Only one enzyme type is utilised in the processes described. There is no description of a method to further enrich the ⁇ -glucan content of the material, or the separation of a distinct layer rich in high molecular weight ⁇ -glucan.
• Lennart et al (US 5,686,123) inform on methods to produce soluble cereal suspensions from oat.
• the basis of the invention is treatment of previously heat-treated ground oat, with ⁇ -amylase class of enzyme, whilst slurried in water.
• a second ⁇ -amylase stage may be optionally included to further breakdown starch.
• No separation of a soluble dietary fibre rich component is described in the invention.
• the product slurry contains most of the protein and oil present in the raw material.
• Triantafyllon in WO 00/24270 describes a method to produce ⁇ -glucan soluble dietary fibre from heat-treated oat flour, using ⁇ -amylase enzyme to hydrolyse starch to lower molecular weight fragments, optionally including ⁇ -amylase and/or protease in a second stage hydrolysis, after which solids are centrifuged off, leaving a single soluble phase containing around up to 2 % ⁇ -glucan before drying.
• ⁇ -amylase enzyme to hydrolyse starch to lower molecular weight fragments, optionally including ⁇ -amylase and/or protease in a second stage hydrolysis, after which solids are centrifuged off, leaving a single soluble phase containing around up to 2 % ⁇ -glucan before drying.
• compositions containing high concentrations of soluble dietary fibres from oat and barley grain are based not on enzymatic extraction, but rather on alkaline extraction either from milled whole grain or a sieved fraction (Fisher et al, US 6,323,338), or even hot water extraction, which yields lower molecular weight soluble ⁇ -glucans (Roxdale Foods Ltd and Morgan; WO 02/02645 Al).
• the invention allows the cost-effective production of oat and barley soluble dietary fibre preparations containing ⁇ -glucans of high molecular weight, in concentrations of typically 20 % - 30 %.
• the fraction containing the high molecular weight soluble dietary fibre component (20% - 30% of dry matter) separates as a distinct viscous top layer during the process, above another distinct aqueous layer containing water soluble components.
• the fraction is relatively free of proteins and oils normally encountered during the processes described above.
• the clean fraction can then be separated very cost effectively from the other components and dries directly as a soluble white powder with negligible cereal taste.
• the main objectives of the present invention are to:
• ⁇ -glucan component of the extract is at least 20 % on a dry matter basis.
• the molecular weight pertains to the demonstrably ⁇ -glucan portion of the complex.
• ⁇ -glucan fraction having a smaller molecular weight such as above 400,000 Daltons.
• the fraction rich in high molecular weight soluble dietary fibre separates from other soluble and water-suspendable components, and from insoluble materials, as a distinct fraction, low in contaminant protein ( ⁇ 7 %) and oil ( ⁇ 2.5 %).
• the mixture can optionally be passed through a wet mill during enzyme treatment, iv. Deactivate the enzymes by heat treatment and allow the hydrolysate mix to settle.
• This sequence crucially facilitates the separation of a distinct fraction such as a settling top layer, in the hydrolysate suspension, which lies above an aqueous layer, with a further distinct bottom layer containing proteins and oils along with the insoluble fibrous portion of the milled grain.
• the top layer is particularly rich in high molecular weight soluble dietary fibre, mainly ⁇ -glucan with some arabinoxylan, alongside maltodextrins and some glucose sugar. This represents a clean separation of a native ⁇ -glucan complex from the other grain components, in that it is believed that the ⁇ -glucan component is close to its original form in the grain.
• ⁇ -glucans In order for the ⁇ -glucans not to become degraded during the enzymatic process, it is essential to start with milled factions of oats that have not been heat-treated and to utilise an amyloglucosidase enzyme preparation that has been cleaned of ⁇ -glucanase side activities. Maintaining the intact ⁇ -glucan structure is a crucial factor in the formation of the distinct top layer, as the separate top layer is not formed if the ⁇ -glucans are degraded.
• This separation is spontaneous in that this soluble dietary fibre rich component separates into a distinct top-layer if the hydrolysate suspension is left without agitation or stirring after completion of the enzymatic stages.
• centrifugation accelerates the formation of this top layer and the use of a 3-phase decanter allows efficient separation of this top-layer from the remainder of the hydrolysate liquor.
• the top-layer When separated, the top-layer contains normally 20 to 30 % (on a dry matter basis), more normally 24 % - 27 %, of high molecular weight ⁇ -glucan, with a low amount of contaminating proteins and oils.
• the layer is readily freeze dried or spray dried to a white - cream coloured powder.
• AMG amyloglucosidase
• AMG amyloglucosidase
• a commercial amyloglucosidase enzyme preparation that has been cleaned of ⁇ -glucanase side activities in a two step procedure using anion exchange, followed by hydrophobic interaction chromatography, the major protein band eluting from the hydrophobic interaction chromatography stage being utilised as the cleaned enzyme.
• the AMG free of ⁇ -glucanase side activities, substantially further degrades dextrins and maltodextrins to low molecular weight oligomers and glucose, whilst leaving the soluble dietary fibre components untransformed/undegraded, facilitating easy separation by ultra-filtration or precipitation into a mix of 50 % ethanol/50 % water, wherein the sugars remain dissolved in the liquid phase and the precipitated, polymeric carbohydrates can be removed by centrifugation, prior to drying.
• a material containing up to 70 % ⁇ -glucan on a dry weight basis, ii.
• the present invention is characterised in that previously non-heat treated de-hulled oat and/or barley grain is first dry milled to an endosperm-starch rich flour fraction and a coarser endosperm-reduced fraction.
• the endosperm-reduced fraction comprises between 45 % and 55 % of the milled grain and is then further utilised without any further heat treatment, which is conventionally applied during oat processing and milling.
• the milled grain is added to water and treated sequentially with a starch degrading ⁇ -amylase enzyme, followed by a second hydrolysis step using an enzyme, or combination of enzymes, from the group amyloglucosidases and pullulanases.
• the enzyme treatments are optionally performed in combination with aqueous wet-milling.
• a further step is enzyme inactivation by wet heat treatment, followed by the spontaneous or centrifugal separation of the hydrolysate mix into a top-layer rich in soluble dietary fibre, mainly ⁇ -glucans, an aqueous layer and a lower layer containing proteins, oils and the insoluble fibrous portion of the grain.
• the present invention relates to a process for the extraction of soluble dietary fibre complex from oat and barley grains using a enzymatic hydrolysis treatment, which is characterized in that the non-heat treated grain is milled and any endosperm depleted fractions thereof being rich in ⁇ -glucans are recombined without any further heat treatment, dispersed in water and then subjected to enzymatic treatment with starch degrading enzymes being free of ⁇ -glucanase, followed by an optional step of enzyme inactivation by wet heat treatment, whereby the hydrolysate mixture forming spontaneously at least one viscous, aqueous top layer upon a second aqueous layer, is subjected to a separation process to isolate said at least one viscous, aqueous top layer comprising the soluble dietary fibre complex, containing more than 20% ⁇ -glucan on a dry matter basis.
• a second aqueous fraction layer substantially free of ⁇ -glucans, and at least a third fraction layer comprising most of the protein and oil together with the insoluble fibrous material from the milled grain are isolated.
• the isolated ⁇ - glucan has a molecular weight of at least 400,000 Daltons.
• the isolated ⁇ -glucan has a molecular weight of at least 800,000 Daltons.
• the isolated ⁇ -glucan has a molecular weight of at least 1,300,000 Daltons.
• the distinct top layer can be removed in a 3-phase decanter or other suitable device, yielding a soluble fraction containing at least 20 % (on a dry matter basis) ⁇ -glucan soluble dietary fibre which is of high molecular weight (> 1,300,000 Daltons) to medium molecular weight (> 800,000 Daltons), along with maltodextrins, arabinoxylans, sugars and relatively low amounts of protein ( ⁇ 7 %) and oils ( ⁇ 2.5 %).
• the separated top layer rich in soluble dietary fibre can then be further treated prior to drying using further enzymatic hydrolysis by way of post-treatment using enzymes of the following types, or combinations of those enzymes: Lichenase, cellulase, xylanase. This allows the reduction of molecular weight of the ⁇ -glucan component of the liquor, and/or the fine tuning of its properties, in a controlled manner.
• the raw material is de-hulled oat, or barley grain, which is dry-milled to remove excess starchy endosperm. Between 45 % - 55 % of the milled grain is retained and used in the wet-process, comprising the coarser fraction. This is not heat treated in the dry state prior to utilisation.
• the milled grain fractions are added to water and then treated with starch-degrading enzymes in a specific sequence, a first stage involving treatment with an enzyme of the amylase type, with optionally concomitant wet-milling, followed by a second stage using an enzyme of the amyloglucosidase and/or pullulanase groups with optionally concomitant wet-milling, whereby the time is up to 40 minutes and treatment at temperatures of 55 °C or greater for the second stage.
• the milled cereal grain fractions are added to water and then treated with starch degrading enzymes in a sequence utilising first ⁇ -amylase and then amyloglucosidase enzyme, in which the amyloglucosidase enzyme is substantially cleaned of ⁇ -glucanase side activities prior to use, in a two step procedure using anion exchange followed by hydrophobic interaction chromatography, the major protein band eluting from the hydrophobic interaction chromatography column being utilised as the cleaned enzyme.
• the hydrolysate spontaneously separates, or is optionally separated centrifugally into 3 distinct layers, a top- layer which is rich in soluble dietary fibres, particularly ⁇ -glucans, but containing little oil ( ⁇ 2.5 %) or protein ( ⁇ 7 %), a middle aqueous layer, and a lower phase containing most of the protein, oil and insoluble fibrous material from the milled grain.
• the top-layer in which the soluble dietary fibres are concentrated is subjected to a further treatment in the wet state using one or a combination of enzymes of the type: Lichenase, cellulase, xylanase. After treatment, the material is heated to inactivate the enzymes and then either freeze dried or spray dried to a powder.
• enzymes of the type Lichenase, cellulase, xylanase.
• the separated top-layer rich in soluble dietary fibre is further treated in the wet state, after optionally further diluting with water, with amyloglucosidase enzyme, in which the amyloglucosidase enzyme is substantially cleaned of ⁇ -glucanase side activities prior to use, in a two step procedure using anion exchange followed by hydrophobic interaction chromatography, isolating the major protein band eluting from the hydrophobic interaction chromatography column and utilising said major protein band as an AMG freed of ⁇ -glucanase side activity as the cleaned enzyme, isolating the fraction rich in ⁇ -glucan and optionally purifying the same from any content of maltose and/or glucose; i.e. by use of ultrafiltration and/or precipitation.
• the isolated ⁇ -glucans can be used in a dry state as well as in a wet state.
• the said fraction contains at least 20 % and up to 40 % ⁇ - glucan soluble dietary fibre, not more than 10 % protein, preferably less than 7 % protein, more preferably less than 5 % protein, and less than 2.5 % oil preferably less than 2.0 %, more preferably less than 1.5 %, still more preferably less than 1.0 %, on a dry matter basis.
• the ⁇ -glucan component has a molecular weight of at least 800,000 Daltons.
• the said fraction contains at least 20 % and up to 40 % ⁇ - glucan soluble dietary fibre, not more than 10 % protein, preferably less than 7 % protein, more preferably less than 5 % protein, and less than 2.5 % oil, preferably less than 2.0 %, more preferably less than 1.5 %, still more preferably less than 1.0 %, on a dry matter basis.
• the ⁇ -glucan component has a molecular weight of at least 1,300,000 Daltons.
• the said fraction contains at least 40 % ⁇ -glucan soluble dietary fibre, not more than 10 % protein, preferably less than 7 % protein, more preferably less than 5 % protein, and less than 2.5 % oil preferably less than 2.0 %, more preferably less than 1.5 %, still more preferably less than 1.0 %, on a dry matter basis.
• the ⁇ -glucan component has a molecular weight of at least 800,000 Daltons.
• the said fraction contains at least 40 % ⁇ -glucan soluble dietary fibre, not more than 10 % protein, preferably less than 7 % protein, more preferably less than 5 % protein, and less than 2.5 % oil preferably less than 2.0 %, more preferably less than 1.5 %, still more preferably less than 1.0 %, on a dry matter basis.
• the ⁇ -glucan component has a molecular weight of at least 1,300,000 Daltons.
• each of the fractions rich in soluble dietary fibre described above is used as an additive for food, feedstuffs, pharmaceuticals, and cosmetics.
• the said fraction which contains at least 40 % ⁇ -glucan soluble dietary fibre, not more than 10 % protein, preferably less than 7 % protein, more preferably less than 5 % protein, and less than 2.5 % oil preferably less than
• the said fraction which contains at least 40 % ⁇ -glucan soluble dietary fibre, not more than 10 % protein, preferably less than 7 % protein, more preferably less than 5 % protein, and less than 2.5 % oil preferably less than 2.0 %, more preferably less than 1.5 %, still more preferably less than 1.0 %, and that has a ⁇ -glucan component with a molecular weight of at least 1,300,000 Daltons, is used as an additive for fruit juice and/or water based drinks.
• a further aspect of the invention relates to the use of the said fraction containing at least 20 % and up to 40 % ⁇ -glucan soluble dietary fibre, not more than 10 % protein, preferably less than 7 % protein, more preferably less than 5 % protein, and less than 2.5 % oil preferably less than 2.0 %, more preferably less than 1.5 %, still more preferably less than 1.0 %, on a dry matter basis, and with the ⁇ -glucan component having a molecular weight of at least 800,000 Daltons, as an additive for yoghurts, milk-based drinks and other liquid fermented milk preparations.
• a further aspect of the invention relates to the use of the said fraction containing at least 20 % and up to 40 % ⁇ -Glucan soluble dietary fibre, not more than 10 % protein, preferably less than 7 % protein, more preferably less than 5 % protein, and less than 2.5 % oil preferably less than 2.0 %, more preferably less than 1.5 %, still more preferably less than 1.0 %, on a dry matter basis, and with the ⁇ -glucan component having a molecular weight of at least 1,300,000 Daltons, as an additive for yoghurts, milk-based drinks and other liquid fermented milk preparations.
• a further aspect of the invention relates to the use of the said fraction containing at least 40 % ⁇ -glucan soluble dietary fibre, not more than 10 % protein, preferably less than 7 % protein, more preferably less than 5 % protein, and less than 2.5 % oil preferably less than 2.0 %, more preferably less than 1.5 %, still more preferably less than 1.0 %, on a dry matter basis, and with the ⁇ -glucan component having a molecular weight of at least 800,000 Daltons, as an additive for yoghurts, milk-based drinks and other liquid fermented milk preparations.
• a further aspect of the invention relates to the use of the said fraction containing at least 40 % ⁇ -glucan soluble dietary fibre, not more than 10 % protein, preferably less than 7 % protein, more preferably less than 5 % protein, and less than 2.5 % oil, preferably less than 2.0 %, more preferably less than 1.5 %, still more preferably less than 1.0 %, on a dry matter basis, and with the ⁇ -glucan component having a molecular weight of not less than 1,300,000 Daltons, as an additive for yoghurts, milk-based drinks and other liquid fermented milk preparations.
• a further aspect of the invention relates to the use of the said fraction containing at least 20 % and up to 40 % ⁇ -glucan soluble dietary fibre, not more than 10 % protein, preferably less than 7 % protein, more preferably less than 5 % protein, and less than 2.5 % oil, preferably less than 2.0 %, more preferably less than 1.5 %, still more preferably less than 1.0 %, on a dry matter basis, and with the ⁇ -glucan component having a molecular weight of at least 800,000 Daltons, as an additive for ice creams and frozen desserts.
• a further aspect of the invention relates to the use of the said fraction containing at least 20 % and up to 40 % ⁇ -glucan soluble dietary fibre, not more than 10 % protein, preferably less than 7 % protein, more preferably less than 5 % protein, and less than 2.5 % oil, preferably less than 2.0 %, more preferably less than 1.5 %, still more preferably less than 1.0 %, on a dry matter basis, and with the ⁇ -glucan component having a molecular weight of at least 1,300,000 Daltons, as an additive for ice creams and frozen desserts.
• a further aspect of the invention relates to the use of the said fraction containing at least 40 % ⁇ -glucan soluble dietary fibre, not more than 10 % protein, preferably less than 7 % protein, more preferably less than 5 % protein, and less than 2.5 % oil, preferably less than 2.0 %, more preferably less than 1.5 %, still more preferably less than 1.0 %, on a dry matter basis, and with the ⁇ -glucan component having a molecular weight of at least 800,000 Daltons, as an additive for ice creams and frozen desserts.
• a further aspect of the invention relates to the use of the said fraction containing at least 40 % ⁇ -glucan soluble dietary fibre, not more than 10 % protein, preferably less than 7 % protein, more preferably less than 5 % protein, and less than 2.5 % oil, preferably less than 2.0 %, more preferably less than 1.5 %, still more preferably less than 1.0 %, on a dry matter basis, and with the ⁇ -glucan component having a molecular weight of at least 1,300,000 Daltons, as an additive for ice creams and frozen desserts.
• a further aspect of the invention relates to the use of the said fraction containing at least 20 % and up to 40 % ⁇ -glucan soluble dietary fibre, not more than 10 % protein, preferably less than 7 % protein, more preferably less than 5 % protein, and less than 2.5 % oil, preferably less than 2.0 %, more preferably less than 1.5 %, still more preferably less than 1.0 %, on a dry matter basis, and with the ⁇ -glucan component having a molecular weight of at least 800,000 Daltons, as an additive for butter based spreads, spreads and margarines, functioning as a blood cholesterol modulating, and /or blood glucose modulating, and/or prebiotic agent.
• a further aspect of the invention relates to the use of the said fraction containing at least 20 % and up to 40 % ⁇ -glucan soluble dietary fibre, not more than 10 % protein, preferably less than 7 % protein, more preferably less than 5 % protein, and less than 2.5 % oil, preferably less than 2.0 %, more preferably less than 1.5 %, still more preferably less than 1.0 %, on a dry matter basis, and with the ⁇ -glucan component having a molecular weight of at least 1,300,000 Daltons, as an additive for butter based spreads, spreads and margarines, functioning as a blood cholesterol modulating, and/or blood glucose modulating, and/or prebiotic agent.
• a further aspect of the invention relates to the use of the said fraction containing at least 40 % ⁇ -glucan soluble dietary fibre, not more than 10 % protein, preferably less than 7 % protein, more preferably less than 5 % protein, and less than 2.5 % oil, preferably less than 2.0 %, more preferably less than 1.5 %, still more preferably less than 1.0 %, on a dry matter basis, and with the ⁇ -glucan component having a molecular weight of at least 800,000 Daltons, as an additive for butter based spreads, spreads and margarines, functioning as a blood cholesterol modulating, and /or blood glucose modulating, and/or prebiotic agent.
• a further aspect of the invention relates to the use of the said fraction containing at least 40 % ⁇ -glucan soluble dietary fibre, not more than 10 % protein, preferably less than 7 % protein, more preferably less than 5 % protein, and less than 2.5 % oil, preferably less than 2.0 %, more preferably less than 1.5 %, still more preferably less than 1.0 %, on a dry matter basis, and with the ⁇ -glucan component having a molecular weight of at least 1,300,000 Daltons, as an additive for butter based spreads, spreads and margarines, functioning as a blood cholesterol modulating, and/or blood glucose modulating, and/or prebiotic agent.
• a further aspect of the invention relates to the use of the said fraction containing at least 20 % and up to 40 % ⁇ -glucan soluble dietary fibre, not more than 10 % protein, preferably less than 7 % protein, more preferably less than 5 % protein, and less than 2.5 % oil, preferably less than 2,0 %, more preferably less than 1.5 %, still more preferably less than 1.0 %, on a dry matter basis, and with the ⁇ -glucan component having a molecular weight of at least 800,000 Daltons, as an additive for cheeses.
• a further aspect of the invention relates to the use of the said fraction containing at least 20 % and up to 40 % ⁇ -glucan soluble dietary fibre, not more than 10 % protein, preferably less than 7 % protein, more preferably less than 5 % protein, and less than 2.5 % oil, preferably less than 2.0 %, more preferably less than 1.5 %, still more preferably less than 1.0 %, on a dry matter basis, and with the ⁇ -glucan component having a molecular weight of at least 1,300,000 Daltons, as an additive for cheeses.
• a further aspect of the invention relates to the use of the said fraction containing at least 40 % ⁇ -glucan soluble dietary fibre, not more than 10 % protein, preferably less than 7 % protein, more preferably less than 5 % protein, and less than 2.5 % oil, preferably less than 2.0 %, more preferably less than 1.5 %, still more preferably less than 1.0 %, on a dry matter basis, and with the ⁇ -glucan component having a molecular weight of at least 800,000 Daltons, as an additive for cheeses.
• a further aspect of the invention relates to the use of the said fraction containing at least 40 % ⁇ -glucan soluble dietary fibre, not more than 10 % protein, preferably less than 7 % protein, more preferably less than 5 % protein, and less than 2.5 % oil, preferably less than 2.0 %, more preferably less than 1.5 %, still more preferably less than 1.0 %, on a dry matter basis, and with the ⁇ -glucan component having a molecular weight of at least 1,300,000 Daltons, as an additive for cheeses.
• a yet further aspect of the invention relates to the use of the said fraction containing at least 20 % and up to 40 % ⁇ -glucan soluble dietary fibre, not more than 10 % protein, preferably less than 7 % protein, more preferably less than 5 % protein, and less than 2.5 % oil, preferably less than 2.0 %, more preferably less than 1.5 %, still more preferably less than 1.0 %, on a dry matter basis, and with the ⁇ -glucan component having a molecular weight of at least 800,000 Daltons, as an additive for processed meat products such as burgers, meatballs, sausages, salamis, pates and pastes, as a texturising and/or moisture retaining agent and/or prebiotic agent and/or blood glucose modulating agent.
• a yet further aspect of the invention relates to the use of the said fraction containing at least 20 % and up to 40 % ⁇ -glucan soluble dietary fibre, not more than 10 % protein, preferably less than 7 % protein, more preferably less than 5 % protein, and less than 2.5 % oil, preferably less than 2.0 %, more preferably less than 1.5 %, still more preferably less than 1.0 %, on a dry matter basis, and with the ⁇ -glucan component having a molecular weight of at least 1,300,000 Daltons, as an additive for processed meat products such as burgers, meatballs, sausages, salamis, pates and pastes, as a texturising and/or moisture retaining agent and/or prebiotic agent and/or blood glucose modulating agent.
• a yet further aspect of the invention relates to the use of the said fraction containing at least 40 % ⁇ -glucan soluble dietary fibre, not more than 10 % protein, preferably less than 7 % protein, more preferably less than 5 % protein, and less than 2.5 % oil, preferably less than 2.0 %, more preferably less than 1.5 %, still more preferably less than 1.0 %, on a dry matter basis, and with the ⁇ -glucan component having a molecular weight of at least 800,000 Daltons, as an additive for processed meat products such as burgers, meatballs, sausages, salamis, pates and pastes, as a texturising and/or moisture retaining agent and/or prebiotic agent and/or blood glucose modulating agent.
• a yet further aspect of the invention relates to the use of the said fraction containing at least 40 % ⁇ -glucan soluble dietary fibre, not more than 10 % protein, preferably less than 7 % protein, more preferably less than 5 % protein, and less than 2.5 % oil, preferably less than 2.0 %, more preferably less than 1.5 %, still more preferably less than 1.0 %, on a dry matter basis, and with the ⁇ -glucan component having a molecular weight of at least 1,300,000 Daltons, as an additive for processed meat products such as burgers, meatballs, sausages, salamis, pates and pastes, as a texturising and/or moisture retaining agent and/or prebiotic agent and/or blood glucose modulating agent.
• a yet further aspect of the invention relates to the use of the said fraction containing at least 20 % and up to 40 % ⁇ -glucan soluble dietary fibre, not more than 10 % protein, preferably less than 7 % protein, more preferably less than 5 % protein, and less than 2.5 % oil, preferably less than 2.0 %, more preferably less than 1.5 %, still more preferably less than 1.0 %, on a dry matter basis, and with the ⁇ -glucan component having a molecular weight of at least 800,000 Daltons, as an additive for baked goods such as breads and cakes as a texturising and/or moisture retaining agent and/or anti- staling agent and/or blood glucose modulating agent and/or a serum cholesterol modulating agent and/or a prebiotic agent.
• a yet further aspect of the invention relates to the use of the said fraction containing at least 20 % and up to 40 % ⁇ -glucan soluble dietary fibre, not more than 10 % protein, preferably less than 7 % protein, more preferably less than 5 % protein, and less than 2.5 % oil, preferably less than 2.0 %, more preferably less than 1.5 %, still more preferably less than 1.0 %, on a dry matter basis, and with the ⁇ -glucan component having a molecular weight of not less than 1,300,000 Daltons, as an additive for baked goods such as breads and cakes as a texturising and/or moisture retaining agent and/or anti-staling agent and/or a blood glucose modulating agent and/or a serum cholesterol modulating agent and/or prebiotic agent.
• a yet further aspect of the invention relates to the use of the said fraction containing at least 40 % ⁇ -glucan soluble dietary fibre, not more than 10 % protein, preferably less than 7 % protein, more preferably less than 5 % protein, and less than 2.5 % oil, preferably less than 2.0 %, more preferably less than 1.5 %, still more preferably less than 1.0 %, on a dry matter basis, and with the ⁇ -glucan component having a molecular weight of at least 800,000 Daltons, as an additive for baked goods such as breads and cakes as a texturising and/or moisture retaining agent and/or anti-staling agent and/or blood glucose modulating agent and/or a serum cholesterol modulating agent and/or a prebiotic agent.
• a yet further aspect of the invention relates to the use of the said fraction containing at least 40 % ⁇ -glucan soluble dietary fibre, not more than 10 % protein, preferably less than 7 % protein, more preferably less than 5 % protein, and less than 2.5 % oil, preferably less than 2.0 %, more preferably less than 1.5 %, still more preferably less than 1.0 %, on a dry matter basis, and with the ⁇ -glucan component having a molecular weight of not less than 1,300,000 Daltons, as an additive for baked goods such as breads and cakes as a texturising and/or moisture retaining agent and/or anti-staling agent and/or a blood glucose modulating agent and/or a serum cholesterol modulating agent and/or prebiotic agent.
• a yet further aspect of the invention relates to the use of the said fraction containing at least 20 % and up to 40 % ⁇ -glucan soluble dietary fibre, not more than 10 % protein, preferably less than 7 % protein, more preferably less than 5 % protein, and less than 2.5 % oil, preferably less than 2.0 %, more preferably less than 1.5 %, still more preferably less than 1.0 %, on a dry matter basis, and with the ⁇ -glucan component having a molecular weight of at least 800,000 Daltons, as a functional additive for cosmetic products such as skin ointments, creams, emollients.
• a yet further aspect of the invention relates to the use of the said fraction containing at least 20 % and up to 40 % ⁇ -glucan soluble dietary fibre, not more than 10 % protein, preferably less than 7 % protein, more preferably less than 5 % protein, and less than 2.5 % oil, preferably less than 2.0 %, more preferably less than 1.5 %, still more preferably less than 1.0 %, on a dry matter basis, and with the ⁇ -glucan component having a molecular weight of at least 1,300,000 Daltons, as a functional additive for cosmetic products such as skin ointments, creams, emollients.
• a yet further aspect of the invention relates to the use of the said fraction containing at least 40 % ⁇ -glucan soluble dietary fibre, not more than 10 % protein, preferably less than 7 % protein, more preferably less than 5 % protein, and less than 2.5 % oil, preferably less than 2.0 %, more preferably less than 1.5 %, still more preferably less than 1.0 %, on a dry matter basis, and with the ⁇ -glucan component having a molecular weight of at least 800,000 Daltons, as a functional additive for cosmetic products such as skin ointments, creams, emollients.
• a yet further aspect of the invention relates to the use of the said fraction containing at least 40 % ⁇ -glucan soluble dietary fibre, not more than 10 % protein, preferably less than 7 % protein, more preferably less than 5 % protein, and less than 2.5 % oil, preferably less than 2.0 %, more preferably less than 1.5 %, still more preferably less than 1.0 %, on a dry matter basis, and with the ⁇ -glucan component having a molecular weight of at least 1,300,000 Daltons, as a functional additive for cosmetic products such as skin ointments, creams, emollients,
• a yet further aspect of the invention relates to the use of the said fraction containing at least 20 % and up to 40 % ⁇ -glucan soluble dietary fibre, not more than 10 % protein, preferably less than 7 % protein, more preferably less than 5 % protein, and less than 2.5 % oil, preferably less than 2.0 %, more preferably less than 1.5 %, still more preferably less than 1.0 %, on a dry matter basis, and with the ⁇ -glucan component having a molecular weight of at least 800,000 Daltons, as a component of a pill or capsule formulation as a prebiotic and/or a blood glucose modulating agent and/or a serum cholesterol modulating agent.
• a yet further aspect of the invention relates to the use of the said fraction containing at least 20 % and up to 40 % ⁇ -glucan soluble dietary fibre, not more than 10 % protein, preferably less than 7 % protein, more preferably less than 5 % protein, and less than 2.5 % oil, preferably less than 2.0 %, more preferably less than 1.5 %, still more preferably less than 1.0 %, on a dry matter basis, and with the ⁇ -glucan component having a molecular weight of at least 1,300,000 Daltons, a component of a pill or capsule formulation as a prebiotic and/or a blood glucose modulating agent and/or a serum cholesterol modulating agent.
• a yet further aspect of the invention relates to the use of the said fraction containing at least 40 % ⁇ -glucan soluble dietary fibre, not more than 10 % protein, preferably less than 7 % protein, more preferably less than 5 % protein, and less than 2.5 % oil, preferably less than 2.0 %, more preferably less than 1.5 %, still more preferably less than 1.0 %, on a dry matter basis, and with the ⁇ -glucan component having a molecular weight of at least 800,000 Daltons, as a component of a pill or capsule formulation as a prebiotic and/or a blood glucose modulating agent and/or a serum cholesterol modulating agent.
• a yet further aspect of the invention relates to the use of the said fraction containing at least 40 % ⁇ -glucan soluble dietary fibre, not more than 10 % protein, preferably less than 7 % protein, more preferably less than 5 % protein, and less than 2.5 % oil, preferably less than 2.0 %, more preferably less than 1.5 %, still more preferably less than 1.0 %, on a dry matter basis, and with the ⁇ -glucan component having a molecular weight of at least 1,300,000 Daltons, a component of a pill or capsule formulation as a prebiotic and/or a blood glucose modulating agent and/or a serum cholesterol modulating agent.
• a yet further aspect of the invention relates to the use of the said fraction containing at least 20 % and up to 40 % ⁇ -glucan soluble dietary fibre, not more than 10 % protein, preferably less than 7 % protein, more preferably less than 5 % protein, and less than 2.5 % oil, preferably less than 2.0 %, more preferably less than 1.5 %, still more preferably less than 1.0 %, on a dry matter basis, and with the ⁇ -glucan component having a molecular weight of at least 800,000 Daltons, as a component in slow and/or controlled released devices for pharmaceutical applications.
• a yet further aspect of the invention relates to the use of the said fraction containing at least 20 % and up to 40 % ⁇ -glucan soluble dietary fibre, not more than 10 % protein, preferably less than 7 % protein, more preferably less than 5 % protein, and less than 2.5 % oil, preferably less than 2.0 %, more preferably less than 1.5 %, still more preferably less than 1.0 %, on a dry matter basis, and with the ⁇ -glucan component having a molecular weight of at least 1,300,000 Daltons, as a component in slow and/or controlled released devices for pharmaceutical applications.
• a yet further aspect of the invention relates to the use of the said fraction containing at least 40 % ⁇ -glucan soluble dietary fibre, not more than 10 % protein, preferably less than 7 % protein, more preferably less than 5 % protein, and less than 2.5 % oil, preferably less than 2.0 %, more preferably less than 1.5 %, still more preferably less than 1.0 %, on a dry matter basis, and with the ⁇ -glucan component having a molecular weight of at least 800,000 Daltons, as a component in slow and/or controlled released devices for pharmaceutical applications.
• a yet further aspect of the invention relates to the use of the said fraction containing at least 40 % ⁇ -glucan soluble dietary fibre, not more than 10 % protein, preferably less than 7 % protein, more preferably less than 5 % protein, and less than 2.5 % oil, preferably less than 2.0 %, more preferably less than 1.5 %, still more preferably less than 1.0 %, on a dry matter basis, and with the ⁇ -glucan component having a molecular weight of at least 1,300,000 Daltons, as a component in slow and/or controlled released devices for pharmaceutical applications.
• Raw material was prepared as follows: Oat grain was first de-hulled and the de-hulled grains were dry milled and 50 % by weight of the grain was retained as a coarser fraction. 575 g of this material was suspended in 4 litres of water at a temperature of 95°C, in a 5 litre reaction vessel fitted with a mechanical stirrer. ⁇ -amylase enzyme (35 units) was added to the suspension and the mixture was incubated, with stirring and intermittent wet-milling, for 1 hour. After this time, the pH was dropped to 4.5, the temperature lowered to 75°C and amyloglucosidase (AMG) enzyme was added (35 units), the mixture being incubated for 15 minutes with stirring. Enzymes were then completely de-activated by heating of the suspension in an autoclave at 140°C for some minutes.
• AMG amyloglucosidase
• the resulting suspension was then centrifuged, producing four distinct layers which were separated and collected: a viscous top layer rich in soluble dietary fibre, particularly ⁇ - glucan, an aqueous layer comprising dextrins and sugars, in particular maltose and maltotriose, ⁇ 1 % fat, and ⁇ 3 % proteins, a protein-oil rich layer and a bottom layer containing the insoluble fibrous part of the milled oat.
• the top-layer and protein-oil layers were freeze dried prior to analysis. The fibrous layer was dried at 60°C in an oven.
• top layer protein-oil fraction and the fibre fraction were 15 %, 15 % and 20.0 % respectively (on a dry matter basis). The remainder was mostly soluble sugars and dextrins.
• the top layer was further analysed for ⁇ -glucan content, residual protein and fat etc with the following results: ⁇ -glucan: 24.5 %, protein: 5.0 %, fat: 1.8 %
• Barley grain was dry milled to remove excess endosperm material and 50 % of the milled grain, representing the coarser fraction, was utilized as the raw material for the trial.
• the resulting suspension was then centrifuged, producing four distinct layers which were separated and collected : a viscous top layer rich in soluble dietary fibre, particularly ⁇ - glucan, an aqueous layer, a protein-oil rich layer and a bottom layer containing the insoluble fibrous part of the milled oat.
• the top-layer and protein-oil layers were freeze dried prior to analysis.
• the fibrous layer was dried at 60°C in an oven.
• top layer The yields of top layer, protein-oil fraction and the fibre fraction were 15 %, 15.4 % and 21.4 % respectively.
• the top layer was further analysed for ⁇ -glucan content, residual protein and etc with the following results: ⁇ -glucan: 24.7 %, protein: 5.1 %, fat: 0.4 %
• a sub-sample of the dried top-layer material was then dissolved in 0.05 M sodium chloride solution to a concentration of 0.1 % and the molecular weight of the polymeric components was estimated using HPSEC on a combined Ultragel GPC column system, using pullulans as Standards.
• the mean peak molecular weight of the ⁇ -glucan component of the material was estimated at > 1.3 million Daltons.
• Example 3 The same raw material as prepared in example 1 was used in this trial. 150 Kg of this material was added to 1050 litres of water at 95°C in a 2,000 litre tank fitted with mechanical stirring. ⁇ -amylase enzyme (9100 units) was added to the suspension and the mixture was incubated, with stirring and intermittent wet-milling, for 1 hour. After this time, the pH was dropped to 4.5 using 84 % orthophosphoric acid, the temperature lowered to 75°C and amyloglucosidase (AMG) enzyme was added (9000 units), the mixture being incubated for 15 minutes with stirring. Enzymes were then completely de-activated by heating the resultant suspension by passing through a tubular heat exchanger at 140°C.
• AMG amyloglucosidase
• the partially cooled hydrolysate suspension was then pumped into a 3-phase decantor and three fractions were obtained: a viscous top-layer rich in soluble dietary fibres, an aqueous fraction and a fraction containing most of the protein, fat and insoluble fibre from the milled oat grain.
• the yields of the top layer, and the protein-oil-fibre fraction were 15.6 % and 35.7 % respectively.
• the separated top layer was then further diluted with water (1 part to 5 parts water), stirred and then excess protein removed centrifugally.
• the cleaned material was then spray dried to a light-cream coloured powder.
• the dried top layer was further analysed for ⁇ -glucan content, sugar, residual protein and oil etc with the following results: ⁇ -glucan: 24.8 %, protein: 5.3 %, fat: 1.6 %
• a sub-sample of the dried top-layer material was then dissolved in 0.05 M sodium chloride solution to a concentration of 0.1 % and the molecular weight of the polymeric components was estimated using HPSEC on a combined Ultragel GPC column system, using pullulans as Standards.
• the mean peak molecular weight of the ⁇ -glucan component of the material was estimated at > 1.3 million Daltons.
• Example 4 A trial equivalent to that described in example 1 was performed with an extra two steps being added to the procedure.
• the separated top-layer was not immediately freeze- dried, but was diluted with water (1 part to 5 parts water) and excess residual protein was removed centrifugally.
• the resulting mix was then passed through an Ultra Filter containing a 0.1 ⁇ m membrane, to remove lower molecular weight components, i.e., sugars such as maltodextrins and glucose.
• the retentate was then collected and freeze- dried. Analysis of the dried fraction gave the following results showed a ⁇ -glucan content of 38.4 % (dry matter basis), with 4.6 % protein.
• GPC analysis of the product after redissolving in 0.05 M sodium chloride solution showed a ⁇ -glucan peak with mean molecular weight of 1,200,500 estimated against pullulan standards.
• Raw material was prepared as follows: Oat grain was first de-hulled and the de-hulled grains were dry milled and 50 % by weight of the grain was retained as the coarser fraction. 575 g of this material was suspended in 4 litres of water at a temperature of 95°C, in a 5 litre reaction vessel fitted with a mechanical stirrer. ⁇ -amylase enzyme (35 units) was added to the suspension and the mixture was incubated, with stirring and intermittent wet-milling, for 1 hour. After this time, the pH was dropped to 5.3, the temperature lowered to 65°C and pullulanase enzyme was added (35 units), the mixture being incubated for 30 minutes with stirring. Enzymes were then completely deactivated by heating of the suspension in an autoclave at 140°C for some minutes.
• the resulting suspension was then centrifuged, producing four distinct layers which were separated and collected : a viscous top layer rich in soluble dietary fibre, particularly ⁇ - glucan, an aqueous layer, a protein-oil rich layer and a bottom layer containing the insoluble fibrous part of the milled oat.
• the top-layer and protein-oil layers were freeze dried prior to analysis.
• the fibrous layer was dried at 60°C in an oven.
• top layer protein-oil fraction and the fibre fraction were 10.3 %, 15.1 % and 15.6 % respectively, on a dry matter basis.
• the top layer was further analysed for ⁇ -glucan content, residual protein and oil etc with the following results: ⁇ -glucan: 18.2 %, protein: 3.9 %, fat: 0.1 %
• a sub-sample of the dried top-layer material was then dissolved in 0.05 M sodium chloride solution to a concentration of 0.1 % and the molecular weight of the polymeric components was estimated using HPSEC on a combined Ultragel GPC column system, using pullulans as Standards.
• the mean molecular weight of the ⁇ -glucan component of the material was estimated at > 1.3 million Daltons.
• the top layer isolated from oat in example 1 was further treated using an amyloglucosidase enzyme preparation which was cleaned of ⁇ -glucanase side activity as follows: 2 ml of AMG was firstly passed through a column containing anion exchange resin (Bio-Rad AG 1-X4) equilibrated in 25 mM phosphate buffer, pH 5.8. Bound protein was then eluted from the column by application of a linear sodium chloride gradient, from 0 to 1 M. The major protein band was collected and re-concentrated to 2 ml using a 1000 Dalton ultrafilter.
• anion exchange resin Bio-Rad AG 1-X4
• the partially cleaned enzyme was then passed onto a column containing hydrophobic interaction chromatography support material (Bio-Rad Macro- Prep t-Butyl HIC Support), equilibrated using 50 mM phosphate buffer, pH 6.0, containing 1.5 M ammonium sulphate. Bound enzyme was then eluted from the column by application of a linear decreasing gradient of ammonium sulphate from 1.5 M to 0. The major protein band eluting from the column was collected, concentrated to 2 ml using a 1000 Dalton ultrafilter and then utilised as cleaned AMG as described below.
• hydrophobic interaction chromatography support material Bio-Rad Macro- Prep t-Butyl HIC Support
• the sample was placed in a water bath at 60°C, with magnetic stirring, and 100 ⁇ l of the cleaned AMG was added to the mix. Incubation was carried out for two hours, after which time the sample was heated to 120°C in an autoclave, to deactivate the enzyme.
• a sub-sample (0.5 ml) was removed from the vessel and was analysed using GPC for the molecular weight distribution of dissolved components, as described in example 1 above.
• the mean molecular weight of the ⁇ -glucan component of the material was measured at > 1.3 million Daltons.
• a peak due to higher molecular weight dextrins encountered in the GPC profile of the product from example 1 had disappeared and a new peak at very low molecular weight was noted, due to dextrin hydrolysis.
• Example 7 A procedure equivalent to that described in example 6 was performed, using the same raw material. However, instead of the hydrolysis product being precipitated after the 2 hour incubation, the liquor was ultra-filtered through an 0.1 ⁇ m membrane, the retentate being subsequently freeze dried.
• ⁇ -amylase enzyme obtained from Genencor
• the aqueous phase was decanted from the solids and analysed. After careful freeze drying, 16.1 g of a cream to light brown powder was obtained which contained about 1.5 g of ⁇ -glucan as determined using the Mcleery enzymatic method. This represents a ⁇ -glucan content of between 9 and 10 % in the separated dry solid.
• the comparative examples show that a separate phase comprising an increased amount of ⁇ -glucans will not be formed.
• the comparative examples show as well as that any ⁇ -glucan will have a very much smaller molecular weight than the ⁇ -glucans isolated according to the present invention.
• Figures 1 and 2 show a schematic overview of the set-up necessary for an industrial process wherein the set-up comprises two parts, viz. a dry process part and a wet process part.
• the dry part (FIG. 1) consists of a bin 1 for storing oat or barley prior to use.
• the grains are transported via a transporting screw 2 to a cleansing means 10 and optionally dosing vessel 3 for weighing in off grains which are transferred to a dehulling apparatus 4, where hulls are taken off via separator 5.
• the dehulled grains are transferred, via a bin 6, to a mill comprising milling rolls and sieves, generally denoted 7, from where flour is retained in a bin 8, and coarser fraction is transferred to and retained in a bin 9 for further treatment.
• the coarser fraction is now transferred to the wet part (FIG. 2) where it is introduced in a reaction vessel 11, together with the enzymes used and water to provide a slurry. pH control sensor (not shown) is applied to the reaction vessel as well as a heating jacket or other temperature controlling means (not shown).
• the reacted mixture is transferred via a wetmill 18 and a heat exchanger 12 to a separator 13 in the form of a decanter, where the top fraction/layer is transferred to a further reaction vessel 14, where the top layer is mixed with water to wash the product by separating of any entrapped protein being removed in a decanter 15, whereupon the ⁇ -glucan fraction is evaporated to produce a ⁇ -glucan powder in driers 16 and 17.
• An intermediate layer is removed as a water phase 19, and a layer comprising solids in the form of fibres, protein and fat is removed as a solids layer 20.

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